Progress Toward the Culture and Transformation of Chicken Blastodermal Cells

AviGenics, Inc., Georgia BioBusiness Center, Athens, 30605, USA.
Stem Cells (Impact Factor: 6.52). 08/2006; 24(7):1638-45. DOI: 10.1634/stemcells.2005-0491
Source: PubMed


Chicken blastodermal cells can be cultured for short periods of time and retain the ability to contribute to somatic and germline tissues when injected into gamma-irradiated stage X embryos. Such a method has yet to yield a germline transgenic bird, in part due to the low rate of transgene integration into the avian genome. In addition, the short culture period precludes the identification and expansion of those cells that carry an integrated transgene. In this study, two methods were developed that produced blastodermal cells isolated from stage X Barred Plymouth Rock embryos bearing an integrated transgene. Addition of chick embryo extract to the culture medium enabled expansion of single colonies for multiple passages. Southern blot analysis indicated that the transgenes had integrated as a single copy in most of the clones. Cells from passaged, transgenic embryo cells were injected into irradiated stage X White Leghorn embryos, producing hatched chicks that bore the donor cells in their somatic tissues. Transgene sequences were detected in sperm DNA; however, breeding of chimeras did not result in germline transmission of the transgene, indicating that the contribution of the transgenic cells to the germline was either nonexistent or very low.

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    • "DNA microinjection into the pronucleus of newly fertilized chicken eggs has been reported since 1994, but so far, it has not been used widely in poultry due to its low transfer efficiency and the laboriousness of the procedure, as described above (Love et al., 1994). In contrast, the liposome transfer method can be applied widely to transfer foreign genes efficiently into cells and embryos (Wang et al., 2006; Suraeva et al., 2008). Never-theless, a recent study on in vitro SSC transfection showed that both the efficiency of transfection and cell survival rate were higher using electroporation rather than liposomes or calcium acid phosphate (20.52 versus 9.75 and 5.61; 69.86 versus 65.00 and 51.16%, respectively) (Yu et al., 2010). "
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